• 대한수의학회지
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• 제36권3호
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• pp.693-698
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• 1996

Haemaphysalis longiscornis is the common cattle tick of great economic importance in Korea. Chemical control using dips or sprays has been the traditional method of attempting to kill these ticks during the infestation period. However, the presence of resistant forms to chemical, the rising costs of acaricides and environmental problems have made it almost impossible to use these chemicals on a regular basis according to the pest problem. For this reason, vaccination against ticks and breeding for host resistance against ticks are being studied. In order to determine the common proteins and antigens according to developmental stages, SDS-PAGE and western blotting were performed. In SDS-PAGE 103.3kD and 98.3kD proteins were observed as common proteins, and these proteins were observed as common antigens in western blotting. Unimmunized rabbits were infestated three times with H longicornis. The weight of the second and the third engorged ticks were 0.153g and 0.104g respectively. This weight is 69% and 47% of the first engorged ticks weight respectively. Immunized rabbits by adult ticks antigen and control were infested with H longicornis. The control taked 3-4 days to fully engorge, but the immunized rabbits taked about 7 days. So adult tick antigen may be effective to render the immunity to host.

• Parasites, Hosts and Diseases
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• 제29권4호
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• pp.339-354
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• 1991

스파르가눔을 흰쥐와 흰생쥐에게 인긍 구강감염을 시켜 감염 2주후부터 숙주의 기생부위에서 충체와 함께 둘레의 조직을 적출하여 동결절편을 만들고 이것을 ABC면역효소표지법과 간접형광항체법 (IFA)으로 항원성분의 분포와 그 추이를 추구한 결과, IgG 항체에 의한 면역반응이 충체의 망상유조직에서 인지되었으며 특히 ABC면역효소표지법에 의하면 유조직 중 수질에서 보다 피질층에서 강한 반응성을 나타났다. 이 중에서도 석회소체가 있는 곳에 염색 반응성이 강하였다. 감염후기에서는 스파르가눔의 외피 (tegument) 내외면과 충체를 둘러 싸고 있는 피낭의 막면, 피막 밖에 있는 섬유성 결합조직과 근조직의 간극에서도 항원성분을 관찰할 수 있다. 특히 결합조직 사이에 분포된 소혈관의 둘레와 현관 외벽에 강한 반응이 나타났다. IgM 항체에 인지된 항원성분도 충체의 유조직층중 피질충에서 강한 반응이 나타났다. 스파르가눔의 추출 조항원성분과 배설분비 항원성분을 SDS-PAGE한 후 nitrocellulose 막에 영동전이하여 IgG, IgM 항체를 유도하는 항원성분을 ABC효소표지 항체 법으로 조사하였다. 스파르가눔의 추출 조항원 성분 중 흰쥐의 IgG 항체에 인지된 항원 성분은 23개이고 IsM 항체에는 15개의 항원성분이 인지되었다. 흰생쥐에서는 IgG 항체로 I6개 항원성분이, IgM 항체에 9개의 항원성분이 인지되었다. 배설분비 항원성분 중 횐쥐의 IgG 항체에 20개 항원성분이, IgM 항체에는 5개의 항원 성분이 인지되었다. 횐생쥐의 IgG 항체에 18개, IgM 항체에 11개의 항원성분이 인지되었다. 조창원성분 중 15개 항원성분과 배설분비 항원성분중 13개의 항원 성분이 흰쥐, 흰생쥐의 IgG 항체에 모두 교차반응이 일어났으며 IgM 항체에 반응한 항원성분중 IgG 항체와도 교차반응이 있었다.

• Parasites, Hosts and Diseases
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• 제40권2호
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• pp.83-88
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• 2002

The 8 kDa antigenic protein of Clonorchis sinensis was partially purified by ammonium sulfate precipitation and subsequently by a column chromatographic steps. The purified protein was separated into 7 and 8 kDa protein bands through SDS-tricine gel electrophoresis, while the protein was fecund to migrate to a 8 kDa band in 7.5-15% SDS-PAGE. The molecular weight of the antigen was estimated to be 110 kDa by Superose 6 HR 10/30 gel filtration. The purified antigen strongly reacted with the human sera of clonorchiasis. The hyperimmune sera of BALB/c mice immunized against the 8 kDa protein were reacted with both the crude extract and the excretory-secretory product of adult worms, but not with the metacercarial extract. Immunohistochemical staining demonstrated that the protein was distributed to the tegument and subtegumental cells and also to the seminal receptacle. The present findings suggest that the 8 kDa protein is a partition of the multicomplek protein originating from various organs of adult C. sinenis, and that it is composed of several 7 and 8 kDa proteins.

• Parasites, Hosts and Diseases
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• 제28권4호
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• pp.207-212
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• 1990

이락촌폐흡충(Paragonimusiloktsuenensis) 감염 흰쥐 혈청의 항체 생산을 효소면역 반응 (enzyme-linked immunosorbent assay: ELISA)으로 관찰하였다. 이락촌폐흡충의 피낭유충을 흰쥐 에 감염시킨 후 격주로 12주까지 채질한 혈청과 이락촌폐흡충 성충 추출 조항원의 반응, 면역교차 반응의 유무를 알기 위해 동혈청과 폐흡충(P. westermani) 조항원(PWA) 및 간흡충(Clonorchis sinensis) 조항원(CSA)의 반응을 ELISA의 OD값(optical density value)으로 측정하였다. 감염강도 별 혈청군(GI, GII, GIII, 및 GIV)은 회수된 충체수로 구분하였다. 이락촌폐흡충 감염 횐쥐 혈청은 동항원(PIA)과의 반응에서 감염 후 4주에 전 감염 개체의 평균 OD값이 대조 혈청과 유의한 차이를 나타내어 항체 양성 반응이 인정되었고, 조사된 12주까지 점차 OD간의 증가로 항체가의 증가를 나타내었다. 감염강도별로 구분한 군별 평균 OD값의 수치는 대체로 감염강도와 비례되었으나 감염 6주 이후 군별 OD값의 유의성은 없었고, 개체별 OD값은 보유충체 수에 비례하지 않는 개체들이 많았다. 이락촌폐흡충 감염 횐쥐 힐청은 폐흡충(p. westermani) 조항원(PWA) 및 간흡충(C. sinensis)조항원(CSA)과의 반응(ELISA)에서도 상당한 면역교차 반응을 나타내었다.

• 한국환경보건학회지
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• 제28권5호
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• pp.28-34
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• 2002

Antigenicity of Rhodotrula rubra isolated from pulmonary tissue of pulmonary tuberculosis patients was studied by means of agglutination reaction with R. rubra whole cell antiserum. And the serological reactivity of crude polyfac charide from R. frubra, Candida albicans, Candida tropicalis, Candida, glabrata, and Saccharomyces cerevisiae ATCC 26603 with antiserum to R. rubra whole cell was studied by means of immunodiffusion test. R. rubra showed stationary phase after 48h when it was cultured in GYEP broth. While agglutinogen titer was 1:64 at lag phase, agglutinogen titer was 1 :256 after 20h. After growth of R. rubra on different 11 media, nutritional environment showed similar agglu-tination reartivity. The agglutinogen titer of C. albicans, C. tropicalis, C. giabrata, which were isolated from patient's expectoration, to R. rubra antiserum by means of agglutination reaction were 1:16, respectively. But, Sacch. cervisiae ATCC26603 was negative. Those results were lower than that of R. rubra agglutinogen titer 1:256. As a result of immu-nodiffusion test with crude polysaccharide extracted from cell wall of R. rubra, C. albicans, C. tropicalis, C. glabrata, Sacch. cervisiae ATCC26603, precipitin line was found only with R. rubra, of which antibody titer was 8.

• 한국식품영양과학회지
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• 제22권1호
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• pp.91-95
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• 1993

Vibrio vulnificus가 생산하는 용혈독소의 단백화학적, 면역화학적 연구에 이용할 목적으로 이 균의 용혈독소에 대한 항혈청을 간편하게 만드는 방법을 실험한 결과는 다음과 같다. Vibrio vulnificus가 생산하는 조용혈독소를 인위적으로 만든 liposome (cholesterol-phos-phatidyl-liposome)에 혼합하여 반응시킨 결과 분자량 50kD의 단백질인 용혈독소만 선택적으로 liposome에 결합되었다. 그러므로 liposome에 결합시킨 조용혈독소를 면역원으로 하고 이 면역원을 토끼의 등근육에 주사하여 항혈청을 간편하게 만들 수 있었으며 이 항혈청은 용혈독소에 대한 특이성이 높았다. 환자와 환경에서 분리된 Vibrio vulnificus의 용혈독소와 liposome를 이용하여 제조한 항혈청을 gel 내 침강반응으로 확인한 결과 단일 침강선을 형성하였다.

• BMB Reports
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• 제38권3호
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• pp.294-299
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• 2005

In this report, we describe the high-yield secretory expression of the recombinant human anti-HBsAg Fab fragment from Pichia pastoris that was achieved by co-integration of the genes encoding the heavy and light chains (both under the control of alcohol oxidase promoter) into the genome of the yeast cells. The fed-batch fermentations were carried out in a 5 L scale. Both chains of the Fab were successfully expressed upon methanol induction. The absorbance ($OD_{600}$) of the broth can reach 350~500 at the end of fed-batch phase. After the induction, the expression level of the recombinant Fab (soluble) reached 420~458 mg/L. The recombinant Fab fragment was purified from the crude culture supernatant by ion exchange chromatography and the purity of the recombinant Fab fragment was over 95%. The affinity activities of the crude fermentation supernatant and the purified Fab were analyzed by indirect ELISA, which showed that the purified recombinant Fab fragment had high affinity activity with hepatitis B surface antigen.

• 한국균학회지
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• 제17권2호
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• pp.66-75
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• 1989

임상가검물(臨床可檢物)로부터 분리(分離)된 Aspergillus fumigatus 균주(菌株)의 항원조성(抗原造成)과 이균에 감염(感染)된 환자의 항체반응(抗體反應)의 다양성을 Immunoblotting으로 관찰하였다. SDS-PAGE와 immunoblots상에 나타난 결과를 보면 비교분석된 배양려액항원(培養慮液抗原)의 조성에 있어서 균주간(菌株間)에 정성(定性) 및 정량적(定量的) 차이가 있음을 알수 있었다. 혈청학적(血淸學的) 반응력(反應力)과 특이성(特異性)이 비교적 높은 항원성분(抗原成分)을 동정(同定)하기 위해 그러한 항원들이 비교적 많이 함유된 AFG7을 선택하였다. 전기영동(電氣泳動)으로 분리하여 nitrocellulose paper로 전이흡착(轉移吸着)시켜 A. fumigatus나 기타 진균항원(眞菌抗原)에 양성침강항체(陽性沈降抗體)(면역확산시험(免疫擴散試驗)에서)를 가진 혈청과 반응시킨 결과 적어도 36개의 항원성분이 검출되었다. 그러나 혈청학적 진단가치가 있는 항원성분은 4개 정도에 불과했다. 특이성과 반응력이 가장 높아 표준항원(標準抗原)에 반드시 함유되어야 할 것으로 보는 항원성분은 분자량(分子量)이 82 KD 정도되는 항원이었고 그 외에 11 KD, 26 KD, 30 KD 및 31 KD 등도 우수한 항원성분으로서 앞으로 더 연구검토되어야 할 것으로 본다. 반응력이 큰 항원이면 Coomassie청(靑)에는 흐리게 염색되어도 immunoblots상에는 짙게 염색되는데 비해 반응력이 약한 항원은 그와 반대현상을 나타내어 immunoblotting 분석법이 항원의 반응력과 특이성 관찰에 매우 유용함을 알 수 있다. 많은 항원성분이 균종특이(菌種特異) 항원결정기(抗原決定基)와 함께 광범한 균군(菌群)에 분포하는 항원결정기(抗原決定基)도 가지고 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제14권11호
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• pp.1598-1604
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• 2001

Eight $12.4{\pm}0.6kg$ initial body weight crossbred barrows were used to determine the effect of soybean galactooligosaccharides on nutrient and energy digestibility, and digesta transit time. Four dietary treatments were utilized in this trial. Treatment one was a corn-soybean meal based diet (SBM) containing raffinose and stachyose at the levels of 0.16% and 0.75%, respectively. Treatment two (control) was a corn-HP300 (soybean concentrate protein) diet. In treatments three and four, 1.1% and 2.2% commercial stachyose was added to the control diet to provide total dietary stachyose at the levels of 1% and 2%, respectively. The soybean galactooligosaccharides (raffinose + stachyose) level in treatment one was slightly lower compared to that in treatment three. Three collection periods were run with two pigs for each treatment/period. There was a 4 d adjustment period followed by a 3 d collection period. The results showed that the nitrogen retention (86.79%) of pigs fed treatment two diet was higher than that of pigs fed treatment one by 5.2% (p<0.05). The nitrogen retention of treatment three was intermediate 83.09%. The apparent fecal digestibility of all amino acids in treatment two was numerically highest, followed by treatments three and four. However, there were no significant difference among groups (p>0.05). The dry matter (DM), organic matter (OM), crude protein (CP), and crude fiber (CF) digestibility numerically decreased as the soybean galactooligosaccharides level increased, but were not significantly different (p>0.05). Chromium content in feces (from the inclusion of 0.3% chromic oxide in the diets) differed among treatments (p<0.05) at 15 h, 18 h, and 21 h after eating. This showed that the digesta transit time was differed significantly among treatments. Treatment four was the shortest, followed by treatment three, SBM and control. The results demonstrated that in the absence of antinutritional factors and soybean antigen protein, inclusion of 1% and 2% stachyose in corn-HP300 diet has no significant effect on the digestibility of DM, OM, CP, CF and amino acids. When the soybean galactooligosaccharide level in diet one and diet three were adjusted to be almost the same, antinutritional factors such as trypsin inhibitor and soybean antigen protein could decrease the nutrient digestibility and nitrogen retention rate of diet. High levels of soybean galactooligosaccharides shortened the digesta transit time in the intestinal tract. This trial suggested that the total level of soybean galactooligosaccharides (stachyose+raffinose) in the weanling piglet diet is better not to exceed 1% when common soybean meal is used as main protein source.

• 대한의생명과학회지
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• 제10권3호
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• pp.219-230
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• 2004

This experiment was performed to examine the in vitro and in vivo affects of the two different haplotype strains of mice, BALB/c and C3H/HeN infected with Echinostoma hortense, and the manifestation of the profiles of cytokine in the splenocytes. In the in vitro experiment, the two mice's splenocytes were divided and stimulated with antigen of crude extracts and the antigen of excretory and secretory products of an adult warm and the manifestation of cytokine mRNA was verified with RT-PCR. As a result, the two different strains of mice both strongly manifested the Th2 cytokine rather than the Thl cytokine and in the case of the Th2 cytokine, the BALB/c mice manifested more strongly than the C3H/HeN mice. In the experiment using the ELISA method, the protem cytokine manifestation had the same result as the mRNA experiment. In the in vivo experiment, the mice was infected via oral route with the metacercaria of the Echinostoma hortense and the manifestation of cytokine was verified by RT-PCR and ELISA and the results were the same as the in vitro experiment. Therefore, in the two strains of BALB/c and C3H/HeN, the C3H/HeN showed a higher susceptivity to the Echinostoma hortense.